Hi Sarona,
FISH combined with immuno very highly depending on the epitope you want to detect thus it may go on chances. However, we do on a regular base consecutive staining with relocalisation. Fist immuno staining and digital imaging then followed by fish and digital imaging. Overlay can be done by software. (for imaging device I can recommend the one from www.3dhistech.com).
For the gene specific probes, especially for custom made set with CE certificate you can use the service from www.kreatech.com they can generate FISH probes for any region of interest.
I hope this may help you. You may contact me for more details.
Karoly
Hi Sarona,
FISH combined with immuno very highly depending on the epitope you want to detect thus it may go on chances. However, we do on a regular base consecutive staining with relocalisation. Fist immuno staining and digital imaging then followed by fish and digital imaging. Overlay can be done by software. (for imaging device I can recommend the one from www.3dhistech.com).
For the gene specific probes, especially for custom made set with CE certificate you can use the service from www.kreatech.com they can generate FISH probes for any region of interest.
I hope this may help you. You may contact me for more details.
Karoly
Thank you very much for the recommendations Karoly,
It is an interesting idea but since I need to demonstrate colocalization/ tight association with a nuclear envelope protein, consecutive staining might not be accurate enough if not imaged simultaneously.
Thanks again,
Sharona
Then the best to try a tyramide signal amplification detection for the proteins before the FISH.
Appl Immunohistochem Mol Morphol. 2008 May;16(3):279-86.
I would use biotin-tyramide system, this leaves bound tyramide at the protein sides, reminaing non-fluorescent and stable enough for consecutive FISH. After the FISH you can detect the biotin using streptavidin in any color you want to combine with the existing FISH colors.
I hope this will be better option for you.
Karoly
The biotin tyramide sounds like a good solution. Worth trying.
Thanks for the useful advice. Sharona
Hi Sharona, You might find this paper of interest: Simultaneous detection of X and Y chromosomes by two-colour
fluorescence in situ hybridization in combination with immunophenotyping of single cells to document chimaerism after
sex-mismatched bone marrow transplantation. MJD van Tol, R Langlois van den Bergh, W Mesker, MCM Ouwerkerk-van Velzen, JM Vossen and HJ Tanke. Bone Marrow Transplantation, (1998) 21, 497–503.
Ann
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